Method for inducing tissue regeneration and treating autoimmune diseases using mesenchymal stem cells

Abstract

The present disclosure relates to a method of inducing tissue regeneration, said method comprising administering to said tissue a composition comprising a population of mesenchymal stem cells (MSCs) and a carrier, wherein said composition possess an inherent ability to reduce inflammation by increasing levels of IL-10 and decreasing levels of TNF-α and IL-6. The present disclosure also relates to a method of treating an autoimmune disease through regeneration of cartilage and bone tissues in a subject, the method comprising: administering therapeutic amount of a composition comprising a population of mesenchymal stem cells (MSCs) and a carrier to the subject by intra-plantar route, intravenous route, intramuscular route, intraarticular route, intraosseous route, or subcutaneous route; wherein said population of MSCs is a homogeneous population having a size in the range of 15-30 μm; and wherein said composition comprises secretome of said MSCs having VEGF and IL-10.

I/We claim: 
     
         1 . A method of inducing tissue regeneration, said method comprising administering to said tissue a composition comprising a population of mesenchymal stem cells (MSCs) and a carrier, wherein at least 70% of the MSCs express at least one marker selected from CD73 CD90 or CD105;
 and less than 5% of MSCs express CD45 marker and HLA-DR marker (Human Leukocyte Antigen—DR isotype); wherein said population of MSCs is a homogeneous population having a size in the range of 15-30 μm; and wherein said composition comprises secretome of said MSCs having VEGF in an amount in the range of 2050 to 2390 μg per million MSCs, and IL-10 in an amount in the range of 1430 to 1690 μg per million MSCs.   
     
     
         2 . The method as claimed in  claim 1 , wherein said tissue is selected from epithelial tissue; connective tissue like bone tissue, cartilage tissue; muscle tissue, elastic tissue, or nervous tissue. 
     
     
         3 . A method of treating an autoimmune disease in a subject in need thereof, the method comprising:
 administering a therapeutically effective amount of a composition comprising a population of mesenchymal stem cells (MSCs) and a carrier to the subject by intra-plantar route, intravenous route, intramuscular route, intraarticular route, intraosseous route, or subcutaneous route;   wherein at least 70% of the MSCs express at least one marker selected from CD73 CD90 or CD105; and less than 5% of MSCs express CD45 marker and (Human Leukocyte Antigen-DR isotype) HLA-DR marker;   wherein said population of MSCs is a homogeneous population having a size in the range of 15-30 μm; and wherein said composition comprises secretome of said MSCs having VEGF in an amount in the range of 2050 to 2390 μg per million MSCs, and IL-10 in an amount in the range of 1430 to 1690 μg per million MSCs.   
     
     
         4 . The method as claimed in  claim 3 , wherein the composition is administered in a range of 1 to 5 doses at a time interval of 1 to 2 months; wherein each dose comprises of 0.5 to 10 million cells per kg of the subject's body weight. 
     
     
         5 . The method as claimed in  claim 3 , wherein the method increases the levels of IL-10 in the subject; wherein the increase in levels of IL-10 estimated 30 days post administration of the composition in the subject having mild autoimmune disease is in the range of 30 to 50% as compared to a control;
 wherein the increase in levels of IL-10 estimated 30 days post administration of the composition in the subject having moderate autoimmune disease is in the range of 40 to 60% as compared to a control;   and wherein the increase in levels of IL-10 estimated 30 days post administration of the composition in the subject having severe autoimmune disease is in the range of 70 to 90% as compared to a control.   
     
     
         6 . The method as claimed in  claim 3 , wherein the method increases the levels of IL-10 in the subject; wherein the increase in levels of IL-10 estimated 60 days post administration of the composition in the subject having moderate autoimmune disease is in the range of 90 to 120% as compared to a control;
 and wherein the increase in levels of IL-10 estimated 60 days post administration of the composition in the subject having severe autoimmune disease is in the range of 80 to 100% as compared to a control.   
     
     
         7 . The method as claimed in  claim 3 , wherein the method decreases the levels of TNF-α in the subject; wherein the decrease in levels of TNF-α estimated in the subject having mild autoimmune disease 30 days post administration of the composition is in the range of 30 to 50% as compared to a control;
 wherein the decrease in levels of TNF-α estimated in the subject having moderate autoimmune disease 30 days post administration of the composition is in the range of 50 to 70% as compared to a control; and 
 wherein the decrease in levels of TNF-α estimated in the subject having severe autoimmune disease 30 days post administration of the composition is in the range of 60 to 80% as compared to a control. 
 
     
     
         8 . The method as claimed in  claim 3 , wherein the method decreases the levels of TNF-α in the subject; wherein the decrease in levels of TNF-α estimated in the subject having moderate autoimmune disease 70 days post administration of the composition is in the range of 80 to 100% as compared to a control; and wherein the decrease in levels of TNF-α estimated in the subject having severe autoimmune disease 70 days post administration of the composition is in the range of 80 to 100% as compared to a control. 
     
     
         9 . The method as claimed in  claim 3 , wherein said subject having mild autoimmune disease exhibits 95 to 100% recovery of symptoms 30 days post administration of the composition; wherein said subject having moderate autoimmune disease exhibits 55 to 65% recovery of symptoms 60 days post administration of the composition; and wherein said subject having severe autoimmune disease exhibits 35 to 45% recovery of symptoms 60 days post administration of the composition. 
     
     
         10 . The method as claimed in  claim 3 , wherein the mild autoimmune disease is mild arthritis with an arthritis scoring in the range of 1≥ to ≤2, and/or a Disease Activity Score in 28 joints (DAS 28 score) in the range of 2.6≥ to ≤3.1; wherein the moderate autoimmune disease is moderate arthritis with an arthritis scoring in the range of 2≥ to ≤3, and/or a DAS 28 score in the range of 3.1≥ to ≤5.1; and wherein the severe autoimmune disease is severe arthritis with an arthritis scoring in the range of 3≥ to ≤4, and/or a DAS 28 score>5.1. 
     
     
         11 . The method as claimed in  claim 1 , wherein said carrier is selected from serelaxin, Ringer's lactate solution, human serum albumin (HSA), DMEM medium, saline solution, phosphate buffered saine (PBS) buffer, Hank's balanced salt solution (HBSS), human plasma, plasma lysate, human albumin, or mixtures thereof, preferably the carrier comprises of serelaxin, Ringer's lactate solution, human serum albumin (HSA), dextran-40, heparin, hyaluronidase or combinations thereof. 
     
     
         12 . The method as claimed in  claim 1 , wherein the MSCs are derived from umbilical cord tissue, cord blood, adipose tissue, bone marrow, or dental pulp, preferably the MSCs are derived from umbilical cord tissue or bone marrow. 
     
     
         13 . The method as claimed in  claim 3 , wherein the MSCs are autogenic or allogenic to the subject. 
     
     
         14 . The method as claimed in  claim 3 , wherein the auto immune disease is selected from the group of consisting of Rheumatoid Arthritis (RA), Acromegaly, Acquired Aplastic Anemia, Acquired Hemophilia, Agammaglobulinemia, Alopecia Areata, Ankylosing Spondylitis (AS), Anti-NMDA Receptor Encephalitis, Antiphospholipid Syndrome (APS), Arteriosclerosis, Autoimmune Addison's Disease (AAD), Autoimmune Autonomic Ganglionopathy (AAG), Autoimmune Encephalitis (AE)/Acute Disseminated Encephalomyelitis (ADEM), Autoimmune Gastritis, Autoimmune Hemolytic Anemia (AIHA), Autoimmune Hepatitis, Autoimmune Hyperlipidemia, Autoimmune Hypophysitis/Lymphocytic Hypophysitis, Autoimmune Inner Ear Disease (AIED), Autoimmune Lymphoproliferative Syndrome (ALPS), Autoimmune Myelofibrosis (AIMF), Autoimmune Myocarditis, Autoimmune Oophoritis, Autoimmune Pancreatitis (AIP), Autoimmune Polyglandular Syndromes (APS), Autoimmune Progesterone Dermatitis (APD), Autoimmune Retinopathy (AIR), Autoimmune Sudden Sensorineural Hearing Loss, Balo Disease/Concentric Sclerosis, Behçet's Disease, Birdshot Chorioretinopathy/Birdshot Uveitis, Bullous Pemphigoid, Castleman Disease, Celiac Disease, Chagas Disease, Chronic Inflammatory Demyelinating Polyneuropathy (CIDP), Chronic Autoimmune Urticaria, Churg-Strauss Syndrome/Eosinophilic Granulomatosis with Polyangiitis (EGPA), Cogan's Syndrome (CS), Cold Agglutinin Disease (CAD), Crest Syndrome, Crohn's Disease, Stricturing Crohn's Disease, Cronkhite-Canada Syndrome (CCS), Cryptogenic Organizing Pneumonia (COP), Dermatitis Herpetiformis (DH), Dermatomyositis, Diabetes, Type 1 (TID), Discoid Lupus Erythematosus (DLE), Dressler's Syndrome/Post myocardial Infarction/Post pericardiotomy Syndrome, Eczema/Atopic Dermatitis, Eosinophilic Fasciitis, Erythema Nodosum, Essential Mixed Cryoglobulinemia, Evans Syndrome, Fibrosing Alveolitis/Idiopathic Pulmonary Fibrosis (IPF), Giant Cell Arteritis/Temporal Arteritis/Horton's Disease, Giant Cell Myocarditis, Glomerulonephritis (GN), Goodpasture's Syndrome/Anti-Gbm/Anti-Tbm Disease, Granulomatosis With Polyangiitis (GPA)/Wegener's Granulomatosis, Graves' Disease (GD), Guillain-Barrè Syndrome (GBS), Hashimoto's Thyroiditis/Autoimmune Thyroiditis, Henoch-Schölein Purpura (HSP)/Iga Vasculitis, Hidradenitis Suppurativa, Hurst's Disease/Acute Hemorrhagic Leukoencephalitis (AHLE), Hypogammaglobulinemia, Iga Nephropathy/Berger's Disease, Immune-Mediated Necrotizing Myopathy (IMNM), Immune Thrombocytopenia (Itp)/Autoimmune Thrombocytopenia Purpura, Inclusion Body Myositis (IBM), Igg4-Related Sclerosing Disease (ISD), Interstitial Cystitis, Juvenile Idiopathic Arthritis (Jia)/Adult-Onset Still's Disease, Juvenile polymyositis//Juvenile dermatomyositis/juvenile myositis, Kawasaki disease, Lambert-Eaton Myasthenic Syndrome (LEMS), Leukocytoclastic vasculitis, Lichen Planus, Lichen Sclerosus, Ligneous conjunctivitis, Linear Iga Disease (LAD), Lupus Nephritis (LN), Lyme Disease/Chronic Lyme Disease/Post-Treatment Lyme Disease Syndrome (PTLDS), Lymphocytic colitis/microscopic colitis, Lymphocytic hypophystitis/autoimmune hypophystitis, Ménière's Disease, Microscopic Polyangiitis (MPA)/ANCA-Associated Vasculitis, Mixed Connective Tissue Disease (MCTD), Mooren's ulcer, Mucha-Habermann disease, Multifocal motor neuropathy, Multiple Sclerosis (MS), Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS), Myasthenia Gravis (MG), Narcolepsy, Neuromyelitis Optica/Devic's Disease, Ocular Cicatricial Pemphigoid, Opsoclonus-myoclonus syndrome (OMS), Palindromic Rheumatism, Paraneoplastic Cerebellar Degeneration (PCD), Paraneoplastic Pemphigus, Parry-Romberg Syndromeherth (PRS)/Hemifacial Atrophy (HFA)/Progressive Facial Hemiatrophy, Paroxysmal Nocturnal Hemoglobinuria (PNH), Peripheral uveitis/pars planitis, PANS/PANDAS, Parsonage-Turner Syndrome (PTS), Pemphigoid Gestationis (PG), Pemphigus Foliaceus, Pemphigus Vulgaris, Pernicious anemia, POEMS Syndrome, Polyarteritis Nodosa (PAN), Polymyalgia Rheumatica, Polymyositis, Postural Orthostatic Tachycardia Syndrome (Pots), Primary Biliary Cirrhosis (PBC), Primary Sclerosing Cholangitis (PSC), Psoriasis, Palmoplantar Pustulosis (PPP), Psoriatic Arthritis, Pulmonary fibrosis, idiopathic (IPF), Pure Red Cell Aplasia (PRCA), Pyoderma gangrenosum, Rasmussen's encephalitis, Raynaud's Syndrome, Reactive Arthritis, Reflex sympathetic dystrophy syndrome (RSD)/Complex regional pain syndrome (CRPS), Relapsing Polychondritis (RP), Restless leg syndrome (RLS)/Willis-Ekbom disease, Rheumatic Fever, Sarcoidosis, Schmidt Syndrome/Autoimmune Polyendocrine Syndrome Type II, Scleritis, Scleroderma, Sclerosing Mesenteritis/Mesenteric Panniculitis, Serpiginous choroidopathy, Sjögren's Syndrome, Stiff person syndrome (SPS), Small Fiber Sensory Neuropathy (SFSN), Small Fiber Sensory Neuropathy (SFSN), Systemic Lupus Erythematosus (SLE), Subacute bacterial endocarditis (SBE), Subacute cutaneous lupus, Susac's syndrome, Sydenham's Chorea, Sympathetic ophthalmia, Takayasu's arteritis (vasculitis), Testicular Autoimmunity, Tolosa-Hunt syndrome, Transverse myelitis (TM), Tubulointerstitial nephritis uveitis syndrome (TINU), Ulcerative Colitis, Undifferentiated Connective Tissue Disease, Uveitis, Vasculitis, VEXAS Syndrome, Vogt-Koyanagi-Harada syndrome (VKH), Osteoarthritis, AVN, vertebral compression factor, urethral stricture, and ureteric stricture.